JP2016507469A - Difluprednate emulsion composition containing antibacterial metal - Google Patents

Abstract

The present invention provides a difluprednate emulsion composition with immediate effect on storage efficacy. The immediate effect of the storage effect is achieved by blending an antibacterial metal (except for zinc). The present invention also provides a method for imparting an immediate effect of preservative effect to an emulsion composition containing difluprednate, comprising preparing an emulsion composition containing difluprednate and an antibacterial metal (excluding zinc). To provide a method characterized by: [Selection figure] None

Description

  The present invention relates to a difluprednate emulsion composition containing an antibacterial metal. More specifically, the present invention relates to a difluprednate-containing emulsion composition having an immediate effect of storage effect by incorporating an antibacterial metal.

(Background of the Invention)
Eye drops, nasal drops, and ear drops are preparations that are used repeatedly, except for those that are used once. The danger of being contaminated with microorganisms is extremely high. Therefore, in order to prevent such secondary contamination, for example, benzalkonium chloride, chlorhexidine gluconate, p-hydroxybenzoate and the like are usually added as preservatives for aqueous preparations. However, emulsions are more difficult to impart storage efficacy than aqueous solutions, and it has been reported that sorbic acid, boric acid and sodium edetate are used as preservatives suitable for emulsions (US Pat. No. 6,379,688). ). The inventors have examined a preservative that can kill bacteria that have invaded in a shorter time than conventional emulsions even when the emulsion is contaminated with secondary microorganisms. In particular, in the preservation efficacy test in the European Pharmacopoeia, it is important to reduce the number of viable bacteria at 6 hours and 24 hours, that is, to kill the invading bacteria in a short time.

  Difluprednate is a potent steroidal anti-inflammatory drug and is marketed in the United States as an emulsion eye drop. This diflupredonate-containing emulsion eye drop contains 0.1 (w / v)% sorbic acid as a preservative.

  On the other hand, a composition containing an antibacterial metal as a preservative has been reported. For example, WO 2004/091567 describes a topical formulation comprising a pigment obtained by stirring a suspension comprising an inorganic pigment and silver oxide, and states that it may be in the form of an emulsion. . WO 2007/012977 describes a foaming composition comprising a steroid, a therapeutically active oil, a surfactant and a polymeric additive, and further describes that silver can be incorporated as an additional therapeutic agent. .

  European Patent No. 0028110 describes a heat stable ophthalmic emulsion composition comprising an aqueous solution of a pharmaceutically acceptable salt of polyacrylic acid crosslinked with triallyl saccharose and describes sulfadiazine silver as an applicable agent. Has been.

  An object of the present invention is to provide an emulsion composition containing difluprednate having an immediate effect of storage efficacy.

  The present inventors have found that a difluprednate-containing emulsion composition having an immediate effect of storage efficacy can be prepared by blending an antibacterial metal.

That is, the present invention is as follows.
[1] An emulsion composition containing difluprednate and an antibacterial metal (excluding zinc).
[2] The emulsion composition according to [1] above, wherein the antibacterial metal (excluding zinc) is a salt or complex of the antibacterial metal (excluding zinc).
[3] The emulsion composition according to [2] above, wherein the salt or complex of the antibacterial metal (excluding zinc) is at least one salt or complex of the antibacterial metal selected from the group consisting of silver and copper .
[4] The emulsion composition according to the above [3], wherein the salt or complex of the antibacterial metal (excluding zinc) is a silver salt or a silver complex.
[5] The emulsion composition according to the above [4], wherein the silver salt or silver complex is at least one selected from the group consisting of silver nitrate, protein silver and silver phytic acid complex.
[6] The emulsion composition according to [4] or [5] above, wherein the concentration of silver salt or silver complex as silver ions is 0.00005 (w / v)% or more and 0.6 (w / v)% or less.
[7] The emulsion composition according to [4] or [5] above, wherein the concentration of silver salt or silver complex as silver ions is 0.0005 (w / v)% or more and 0.01 (w / v)% or less.
[8] The emulsion composition according to the above [3], wherein the salt or complex of the antibacterial metal (excluding zinc) is a copper salt or a copper complex.
[9] The emulsion composition according to the above [8], wherein the copper salt or copper complex is copper sulfate.
[10] The emulsion composition according to the above [8] or [9], wherein the concentration of the copper salt or copper complex as a copper ion is higher than 0.0001 (w / v)% and not higher than 0.5 (w / v)%.
[11] The emulsion composition according to the above [8] or [9], wherein the concentration of the copper salt or copper complex as a copper ion is 0.0005 (w / v)% or more and 0.01 (w / v)% or less.
[12] The emulsion composition according to any one of [1] to [11] above, which is an ophthalmic composition.
[13] A method for imparting an immediate effect of storage efficacy to an emulsion composition containing difluprednate, comprising preparing an emulsion composition containing difluprednate and an antibacterial metal (excluding zinc) A method characterized by.
[14] The method according to [13] above, wherein the immediate effect of the storage effect is to reduce the viable cell count by 3 logs or more within 24 hours after inoculation.
[15] The immediate effect of the preservation effect is to reduce the number of viable bacteria by 3 logs or more within 24 hours after inoculation and to reduce the number of viable bacteria by 2 logs or more within 6 hours after inoculation [ [13] The method described.
[16] The method according to any one of [13] to [15] above, wherein the antibacterial metal (excluding zinc) is a salt or complex of the antibacterial metal (excluding zinc).
[17] The method according to [16] above, wherein the salt or complex of the antibacterial metal (excluding zinc) is at least one salt or complex of the antibacterial metal selected from the group consisting of silver and copper.
[18] The method according to [17] above, wherein the salt or complex of the antibacterial metal (excluding zinc) is a silver salt or a silver complex.
[19] The method described in [18] above, wherein the silver salt or silver complex is at least one selected from the group consisting of silver nitrate, protein silver and silver phytic acid complex.
[20] The method according to [18] or [19] above, wherein the silver salt or silver complex is blended as silver ions at a concentration of 0.00005 (w / v)% or more and 0.6 (w / v)% or less.
[21] The method according to [18] or [19] above, wherein the silver salt or silver complex is blended as silver ions at a concentration of 0.0005 (w / v)% or more and 0.01 (w / v)% or less.
[22] The method according to [17] above, wherein the salt or complex of the antibacterial metal (excluding zinc) is a copper salt or a copper complex.
[23] The method described in [22] above, wherein the copper salt or copper complex is copper sulfate.
[24] The method according to [22] or [23] above, wherein the copper salt or copper complex is blended at a concentration of higher than 0.0001 (w / v)% and lower than or equal to 0.5 (w / v)% as copper ions.
[25] The method according to [22] or [23] above, wherein the copper salt or copper complex is blended as a copper ion at a concentration of 0.0005 (w / v)% or more and 0.01 (w / v)% or less.
[26] The method according to any one of [13] to [25] above, wherein the emulsion composition is an ophthalmic composition.
[27] A method for imparting an immediate effect of storage efficacy to an emulsion composition containing difluprednate by blending an antibacterial metal (excluding zinc) with the emulsion composition containing difluprednate.
[28] The method according to [27] above, wherein the immediate effect of the preservation effect is to reduce the viable cell count by 3 logs or more within 24 hours after inoculation.
[29] The immediate effect of the preservation effect is to reduce the number of viable bacteria by 3 logs or more within 24 hours after inoculation and to reduce the number of viable bacteria by 2 logs or more within 6 hours after inoculation. 27] The method described.

(Detailed description of the invention)
The present invention provides an emulsion composition containing difluprednate and an antibacterial metal (except zinc). More specifically, the present invention relates to an oil-in-water emulsion composition (hereinafter referred to as the present invention) containing difluprednate, oil, water, an emulsifier and an antibacterial metal (excluding zinc), which has an immediate effect of storage effect. The composition of the invention).

  In this specification, unless otherwise specified, having an immediate effect of storage efficacy means, for example, reducing the number of viable bacteria in a short time in a storage efficacy test, for example, bacteria (S. aureus, E. coli). And P. aeruginosa) can be reduced by 3 logs or more within 24 hours after inoculation. Furthermore, it means that the number of viable bacteria can be reduced by 3 logs or more within 24 hours after inoculation and the number of viable bacteria can be reduced by 2 logs or more within 6 hours after inoculation. The above-mentioned standard is one of the judgment standards when the preservation efficacy test described in European Pharmacopoeia 7.0 (EUROPEAN PHARMACOPOEIA 7.0, Efficacy of antimicrobial preservation) is performed.

  The operation method of the preservation efficacy test described in European Pharmacopoeia 7.0 uses bacteria, Staphylococcus aureus and Pseudomonas aeruginosa, and fungi Candida albicans and In this method, black mold (Aspergillus brasiliensis (niger)) is used and the following operations (i) to (iv) are performed. If necessary, bacteria such as Escherichia coli can be added to the test bacteria.

  (i) Each of the above five strains used in the test is inoculated on the surface of an agar slant medium and precultured. As the agar medium for preculture, a soy bean / casein / digest agar medium is used in the case of bacteria, and a Sabouraud dextrose agar medium is used in the case of fungi. Bacteria for 18-24 hours at 30-35 ° C, Candida albicans for 40-48 hours at 20-25 ° C, Aspergillus brasiliensis (niger) at 20-25 ° C for 1 week or until sufficient spores are formed Incubate.

(ii) Using the aqueous liquid composition to be tested as a sample, dispense 10 mL of each sample into 5 sterilized test tubes with stoppers, and transfer the test bacteria in (i) to 10 ⁵ to 10 ⁶ cells / After preparing the mixed sample inoculated to be mL, store it in the dark at 20-25 ° C. In addition, test bacteria are not mixed, but inoculate each specimen individually.

  (iii) After storage for 24 hours from the start of storage, sample 1 mL from each mixed sample and dilute the solution with 9 mL of physiological saline. Further, perform the same dilution 2-3 times, and dispense 1 mL of each diluted solution into a sterile petri dish.

(iv) Next, lecithin 0.1 (w / v)% polysorbate 80 0.7 (w / v)% added soybean casein digest agar medium for fungi, lecithin 0.1 (w / v)% polysorbate 80 0.7 ( After adding w / v)% added Sabouraud-glucose agar medium and culturing under the following conditions, the number of generated colonies is measured, and the theoretical number of bacteria per 1 mL of the mixed sample is calculated.
Bacterial culture conditions: about 3 days at 30-35 ° C Fungal culture conditions: about 5 days at 20-25 ° C

  After the above operations (i) to (iv), when the viable count of all the bacteria (S. aureus, Escherichia coli and Pseudomonas aeruginosa) in the mixed solution is reduced by 3 logs or more within 24 hours after inoculation, Judged to have immediate effect on storage efficacy. Furthermore, when the number of viable bacteria is reduced by 3 logs or more within 24 hours after inoculation and the number of viable bacteria is reduced by 2 logs or more within 6 hours after inoculation, it is determined to have an immediate effect of preservation effect.

  Difluprednate (6α, 9α-difluoroprednisolone 17-butyrate 21-acetate) that can be used in the composition of the present invention is a steroidal anti-inflammatory drug, and has an excellent anti-inflammatory effect by transdermal administration or eye drop administration. And is known to exhibit antiallergic activity. Difluprednate can be prepared, for example, based on the methods described in US Pat. No. 3,780,177 and US Pat. No. 3,786,492.

  The oil that can be used in the composition of the present invention is not particularly limited as long as it can be applied to eyes with low toxicity and low irritation, and preferably contains a fatty acid ester of glycerin, such as castor oil, peanut oil, cottonseed oil. , Soybean oil, olive oil, medium chain fatty acid triglycerides [for example, miglyol (trade name, Mitsuba Trade)] and the like. More preferable examples include castor oil and medium chain fatty acid triglyceride (for example, miglyol) which can dissolve difluprednate well, and particularly preferable is castor oil.

  The water that can be used in the composition of the present invention is not particularly limited as long as it is usually added to a pharmaceutical composition, and examples thereof include purified water and distilled water for injection.

  Examples of emulsifiers that can be used in the composition of the present invention include nonionic surfactants. Examples thereof include polyoxyethylene sorbitan fatty acid esters, polyoxyethylene hydrogenated castor oil, alkylaryl polyether alcohol type polymers, polyoxyethylene fatty acid esters, polyoxyethylene polyoxypropylene glycol or sucrose fatty acid esters, preferably Polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene sorbitan monostearate, polyoxyethylene sorbitan tristearate, polyoxyethylene hydrogenated castor oil 10, polyoxy Ethylene hardened castor oil 40, polyoxyethylene hardened castor oil 50, polyoxyethylene hardened castor oil 60, tyloxapol, polyoxyl stearates And the like, particularly preferably polysorbate 80, polyoxyethylene hydrogenated castor oil 60, a tyloxapol and polyoxyl 40 stearate. Further, these may be used in combination.

  The antibacterial metal (excluding zinc) in the composition of the present invention is preferably a compound in which the antibacterial metal (excluding zinc) can be an ion in the emulsion composition. It may be in the form of a salt or complex of an antibacterial metal (excluding zinc). The antibacterial metal (excluding zinc) in the composition of the present invention may be any antibacterial metal (excluding zinc), such as silver or copper, or a combination thereof. . Silver or copper is preferably a compound that can become silver ions or copper ions in the emulsion composition.

  Examples of compounds that can be silver ions include metallic silver, silver salts, silver complexes, and the like, and silver salts or silver complexes are preferable.

  The metallic silver may be nanoparticulate silver or colloidal silver.

  Examples of silver salts include silver nitrate, silver sulfate, silver chloride, silver bromide, silver oxide, silver acetate, silver carbonate, silver citrate, silver lactate, silver phosphate, silver oxalate, silver thiosulfate, and silver protein. Preferred are silver nitrate and protein silver.

  Known silver complexes include silver cyanide complex, diamine silver complex, silver thiosulfate complex, tetrakis (pyridine) silver (II) peroxodisulfate, silver chloro complex salt, silver amino acid complex, silver phytic acid complex, and the like. . A silver phytic acid complex is preferred. For example, LunarSilver (registered trademark) (containing 0.5 (w / v)% silver and 1.5 (w / v)% phytic acid) can be used as the silver phytic acid complex. Here, LunarSilver (registered trademark) is obtained by stabilizing silver in an aqueous solution manufactured and sold by Antibacterial Chemical Research Co., Ltd. with phytic acid.

  Examples of compounds that can be copper ions include metal copper, copper salts, copper complexes, and the like, and copper salts and copper complexes are preferable.

  Metallic copper may be nanoparticulated copper or colloidal copper.

  Examples of the copper salt include copper sulfate, copper chloride, copper oxide, and copper carbonate. Copper sulfate is preferred.

  Copper complexes include copper-ethanolamine, copper-dimethyldithiocarbamate, copper-sulfate, copper-2-ethylhexanoate, copper-quaternary alkylammonium, copper hydroxide (II), basic copper carbonate, copper -Nitrate, copper-8-quinolinolate, copper amino acid complex and the like.

  The blending ratio of difluprednate in the composition of the present invention is, for example, 0.001 (w / v)% or more, preferably 0.005 (w / v)% or more, more preferably 0.01 (w / v) in the composition. % Or more, 0.4 (w / v)% or less, preferably 0.3 (w / v)% or less, more preferably 0.2 (w / v)% or less.

  The blending ratio of oil in the composition of the present invention is not particularly limited as long as it is usually a blending ratio capable of preparing an oil-in-water emulsion, and the blending ratio is 0.1 (w / v)% or more in the composition. , Preferably 0.5 (w / v)% or more, more preferably 1 (w / v)% or more, 40 (w / v)% or less, preferably 30 (w / v)% or less, more preferably 20 (w / v)% or less.

  The mixing ratio of water in the composition of the present invention is not particularly limited, but is 20 (w / v)% or more, preferably 50 (w / v)% or more, more preferably 60 (w / v)% in the composition. These are 99.8 (w / v)% or less, preferably 99 (w / v)% or less, more preferably 98 (w / v)% or less.

  The blending ratio of the emulsifier in the composition of the present invention is not particularly limited, and may be any blending ratio that can usually prepare an oil-in-water emulsion, and the blending ratio is 0.1 (w / v)% or more in the composition. , Preferably 0.5 (w / v)% or more, more preferably 1 (w / v)% or more, 40 (w / v)% or less, preferably 30 (w / v)% or less, more preferably 20 (w / v)% or less.

  The proportion of the antibacterial metal (except zinc) in the composition of the present invention is usually about 0.00005 (w / v)% or more and about 0.6 (in terms of silver ion) in the case of a silver salt or a silver complex. w / v)% or less, preferably about 0.0001 (w / v)% or more and about 0.01 (w / v)% or less, more preferably about 0.0005 (w / v)% or more and about 0.01 ( w / v)% or less, and most preferably about 0.005 (w / v)% or more and about 0.01 (w / v)% or less. From the viewpoint of changing the appearance of the composition, the upper limit of addition as silver ions is preferably about 0.01 (w / v)%. In the case of a copper salt or a copper complex, the concentration as a copper ion is usually higher than about 0.0001 (w / v)% and about 0.5 (w / v)% or less, preferably about 0.0005 (w / v)% or more. , About 0.01 (w / v)% or less. From the viewpoint of eye safety, the upper limit of addition as copper ions is preferably about 0.01 (w / v)%.

  The combination of the above-mentioned components in the composition of the present invention is not particularly limited.For example, in the composition, difluprednate 0.001 to 0.4 (w / v)%, oil 0.1 to 40 (w / v)%, 20 ~ 99.8 (w / v)% of water, 0.1 ~ 40 (w / v)% of emulsifier, and antibacterial metal (except for zinc) is silver salt or silver complex, the concentration as silver ion is about 0.00005 (w / v)% or more, about 0.6 (w / v)% or less, and in the case of copper salt or copper complex, the concentration as copper ion is higher than about 0.0001 (w / v)% and about 0.5 (w / v) )% Or less. Preferably in the composition, difluprednate 0.005-0.3 (w / v)%, oil 0.5-30 (w / v)%, water 50-99 (w / v)%, emulsifier 0.5-30 (w / v )%, And when the antibacterial metal (excluding zinc) is a silver salt or a silver complex, the concentration as silver ions is about 0.0005 (w / v)% or more, about 0.01 (w / v)% or less, In the case of a copper salt or a copper complex, the concentration as a copper ion is blended at a ratio of about 0.0005 (w / v)% or more and about 0.01 (w / v)% or less. Particularly preferably in the composition, difluprednate 0.01-0.2 (w / v)%, oil 1-20 (w / v)%, water 60-98 (w / v)%, emulsifier 1-20 (w / v). v)% and antibacterial metals (excluding zinc), in the case of silver salts or silver complexes, the concentration as silver ions is about 0.005 (w / v)% or more and about 0.01 (w / v)% or less In the case of a copper salt or a copper complex, the concentration as a copper ion is blended at a ratio of about 0.0005 (w / v)% or more and about 0.01 (w / v)% or less. The most preferred composition of the present invention contains difluprednate 0.05 (w / v)%, castor oil 5.0 (w / v)%, polysorbate 80 4.0 (w / v)% in each component, and antibacterial In the case of silver salts or silver complexes, the concentration as silver ions is about 0.00005 to about 0.0005 (w / v)%, and the concentration as copper ions in the case of copper salts or copper complexes Contains about 0.0005 (w / v)% of each component and has a pH of about 5.5.

  In the composition of the present invention, a water-soluble polymer can be blended in order to increase the stability of the emulsion particles. Examples of the water-soluble polymer include povidone (polyvinyl pyrrolidone), polyvinyl alcohol, hydroxyethyl cellulose, hydroxypropyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, carboxymethyl cellulose, carboxyvinyl polymer, and salts thereof. The water-soluble polymer can be added to the composition in an amount of about 0.001 to about 3 (w / v)%.

  An isotonic agent can be mix | blended with the composition of this invention. Examples of tonicity agents include boric acid, sodium chloride, potassium chloride, concentrated glycerin, propylene glycol, D-mannitol and the like. The above-mentioned isotonic agents can be added as long as the storage stability of the difluprednate is not significantly reduced and the physical stability of the emulsion is not impaired. In particular, any of boric acid, sodium chloride and concentrated glycerin, which hardly affects the storage stability of difluprednate, is preferred. Further, these may be used in combination.

  The composition of the present invention is prepared at an osmotic pressure of about 150 to about 1100 mOsm, preferably at an osmotic pressure of about 150 to about 650 mOsm, and more preferably by the addition of an isotonic agent as described above. The osmotic pressure is prepared at about 220 to about 480 mOsm.

  A buffering agent can be mix | blended with the composition of this invention. Examples of buffers include acetates such as sodium acetate, phosphates such as monosodium dihydrogen phosphate, disodium monohydrogen phosphate, dipotassium dihydrogen phosphate, dipotassium monohydrogen phosphate, and epsilon aminocaproic acid. And amino acid salts such as sodium glutamate, citric acid and its salts, trometamol, 4- (2-hydroxyethyl) -1-piperazine ethanesulfonic acid and the like.

  A buffering agent can be added as long as the storage stability of the difluprednate is not lowered and the physical stability of the emulsion is not impaired. Buffering agents can be added to the composition from about 0.01 to about 2 (w / v)%.

  Anti-inflammatory agents such as anti-inflammatory agents, non-steroidal anti-inflammatory agents, anti-inflammatory analgesics, anti-inflammatory enzyme agents, antiviral agents, antibacterial agents, antifungal agents, antiallergic agents, antibiotics, sulfa drugs, synthetic Penicillin, glaucoma treatment, cataract treatment, miosis, mydriasis, local astringent, vasoconstrictor, intraocular hypertension, ocular hypertension, surface anesthetic, α1-blocker, β-blocker Agents, β1-blockers, carbonic acid dehydrogenase inhibitors, selective H1-blockers for topical use, corticosteroids, vitamin B12, coenzyme-type vitamin B2, anticholinesterase agents, organic iodine preparations, and the like can also be added.

  In addition to the composition of the present invention, various additives such as a stabilizer, an antioxidant, a chelating agent, a pH adjuster, and a thickener can be blended. Further, a preservative other than the antibacterial metal may be further added. Examples of the antioxidant include ascorbic acid and its salt, tocopherol, sodium thiosulfate, sodium bisulfite, pyruvic acid and its salt, and the like. Examples of chelating agents include sodium edetate, citric acid and salts thereof. Examples of the pH adjuster include hydrochloric acid, phosphoric acid, acetic acid, sulfuric acid, boric acid, borax, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, aqueous ammonia and the like. Examples of particularly acidic pH adjusting agents include hydrochloric acid, phosphoric acid, acetic acid, sulfuric acid, and boric acid. Examples of the stabilizer include dibutylhydroxytoluene, trometamol, sodium formaldehyde sulfoxylate, tocopherol, sodium pyrosulfite, monoethanolamine, aluminum monostearate, glyceryl monostearate and the like. Examples of the thickener include carboxyvinyl polymer, hydroxyethyl cellulose, hydroxypropylmethyl cellulose, methyl cellulose, alginic acid, polyvinyl alcohol, polyvinyl pyrrolidone, macrogol, sodium hyaluronate and the like. Examples of preservatives that may be added in addition to antibacterial metals (except zinc) include benzalkonium chloride, benzethonium chloride, chlorhexidine gluconate, chlorobutanol, sorbic acid, potassium sorbate, sodium dehydroacetate And methyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, butyl paraoxybenzoate and the like.

  The composition of the present invention can be provided as an aqueous preparation such as an oil-in-water (O / W type) emulsion or microemulsion.

  The average particle diameter (median diameter) of the oil droplets of the composition of the present invention is preferably 10 to 2000 nm, more preferably 20 to 1000 nm, and particularly preferably 20 to 500 nm. The average particle size can be measured using a particle size distribution measuring apparatus.

  The pH of the composition of the present invention is preferably 3-8. A more preferred pH is 4-7, a further preferred pH is 5-6, and a most preferred pH is about 5.5. In this pH range, the stability of difluprednate is best.

  The composition of the present invention is prepared by preparing an aqueous phase containing an emulsifier and an oil phase containing difluprednate, and mixing and emulsifying them. In order to uniformly emulsify, known means such as a homomixer, a homogenizer, a high-pressure homogenizer, and an ultrahigh-pressure homogenizer (microfluidizer) can be used. Other additives such as antibacterial metals (excluding zinc), isotonic agents and buffering agents may be dissolved in an aqueous phase containing an emulsifier, or may be added to an emulsified emulsion.

  In particular, a process of preparing an aqueous phase by adding an antibacterial metal (except zinc), an emulsifier, an isotonic agent and a buffer to water, dissolving a difluprednate in oil, and preparing a difluprednate oil phase A step of mixing the aqueous phase and the difluprednate oil phase, roughly emulsifying with a homogenizer or the like to prepare a coarse emulsion, and further finely pulverizing the coarse emulsion with a homogenizer to form an oil-in-water emulsion. It is desirable to manufacture from the process of preparing. In order to suppress a decrease in the stability of difluprednate during the production process, the aqueous phase is adjusted to pH 5-6 by adding a pH adjusting agent before mixing with the difluprednate oil phase. It is desirable to do.

  In the present specification, “emulsification” refers to making the oil phase into a large number of fine liquid droplets and dispersing and holding them in the water phase. “Coarse emulsification” refers to a state in which the oil phase is made into fine droplets to some extent and dispersed and held in the aqueous phase in one state of emulsification. At this time, the droplet size is non-uniform. “Particulate formation” refers to a state of emulsification, in which a crude emulsion is further finely divided into droplets of oil phase by using a device such as a microfluidizer, and the droplet size is made uniform to some extent. .

  In addition, in the preparation process of the composition of the present invention, additives such as antibacterial metals (excluding zinc), tonicity agents, buffering agents, etc. may be dissolved in the aqueous phase, or the emulsion after emulsification It may be added.

  The composition of the present invention is preferably used as a preparation for topical administration to the eyes, nose, ears or skin, and more preferably used as an ophthalmic composition such as eye drops, nasal drops, ear drops or lotions. .

  The composition of the present invention has excellent anti-inflammatory action, anti-allergic action and antibacterial action. Therefore, it is useful for the prevention and treatment of various inflammatory diseases such as allergic conjunctivitis, spring catarrh, blepharitis, catarrhal conjunctivitis, uveitis, inflammation or pain associated with eye surgery, and macular edema. It can also be used advantageously when administered locally to the eyes, nose, ears, skin, and the like.

  The composition of the present invention can be safely administered to mammals (human, dog, rabbit, cow, horse, monkey, cat, sheep, etc.).

  The dose of the composition of the present invention varies depending on the type of disease, symptoms, patient age, weight, etc. For example, when used as an ophthalmic solution for adults, 0.01 mg of difluprednate per eye per patient is used. As an ophthalmic solution containing ˜0.2 (w / v)%, it is desirable to administer 1 to 2 drops at a time, about 2 to 4 times a day, depending on the symptoms.

  In addition, the present invention is a method for imparting an immediate effect of storage efficacy to an emulsion composition containing difluprednate, and prepares an emulsion composition containing difluprednate and an antibacterial metal (excluding zinc). To provide a method characterized by: More specifically, (a) difluprednate, (b) water, (c) oil, (d) emulsifier and (e) antibacterial metal (except zinc) are mixed to form an oil-in-water emulsion composition. A method for imparting an immediate effect of storage efficacy to an emulsion composition containing difluprednate (hereinafter referred to as the method of the present invention).

  In the method of the present invention, to give an immediate effect of the preservation effect is, for example, to reduce the number of viable bacteria by 3 logs or more within 24 hours after inoculation, and further to reduce the number of viable bacteria within 24 hours after inoculation. It is to reduce 3 log or more and to reduce the number of viable bacteria by 2 log or more within 6 hours after inoculation.

  The difluprednate that can be used in the method of the present invention is as described above.

  Examples of the oil that can be used in the method of the present invention include castor oil, peanut oil, cottonseed oil, soybean oil, olive oil, medium chain fatty acid triglycerides [for example, miglyol (trade name, Mitsuba Trade)] and the like. Can be mentioned. More preferable are castor oil and medium chain fatty acid triglyceride (for example, miglyol) having high solubility of difluprednate, and particularly preferable is castor oil.

  Examples of the type of emulsifier that can be used in the method of the present invention include nonionic surfactants as described above. Examples thereof include polyoxyethylene sorbitan fatty acid esters, polyoxyethylene hydrogenated castor oil, alkylaryl polyether alcohol type polymers, polyoxyethylene fatty acid esters, polyoxyethylene polyoxypropylene glycol or sucrose fatty acid esters, preferably Polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene sorbitan monostearate, polyoxyethylene sorbitan tristearate, polyoxyethylene hydrogenated castor oil 10, polyoxy Ethylene hardened castor oil 40, polyoxyethylene hardened castor oil 50, polyoxyethylene hardened castor oil 60, tyloxapol, polyoxyl stearates And the like, particularly preferably polysorbate 80, polyoxyethylene hydrogenated castor oil 60, a tyloxapol and polyoxyl 40 stearate. Further, these may be used in combination.

  The antibacterial metal (excluding zinc) in the method of the present invention is preferably a compound in which the antibacterial metal (excluding zinc) can be an ion in the emulsion composition. The antibacterial metal (except for zinc) may be in the form of a salt or complex. The antibacterial metal (excluding zinc) in the composition of the present invention may be any metal having antibacterial properties, such as silver or copper, or a combination thereof. Silver or copper is preferably a compound that can become silver ions or copper ions in the emulsion composition.

  Examples of compounds that can be silver ions include metallic silver, silver salts, silver complexes, and the like, and silver salts or silver complexes are preferable.

  The metallic silver may be nanoparticulate silver or colloidal silver.

  Examples of silver salts include silver nitrate, silver sulfate, silver chloride, silver bromide, silver oxide, silver acetate, silver carbonate, silver citrate, silver lactate, silver phosphate, silver oxalate, silver thiosulfate, and silver protein. Preferred are silver nitrate and protein silver.

  Known silver complexes include silver cyanide complex, diamine silver complex, silver thiosulfate complex, tetrakis (pyridine) silver (II) peroxodisulfate, silver chloro complex salt, silver amino acid complex, silver phytic acid complex, and the like. . A silver phytic acid complex is preferred. For example, LunarSilver (registered trademark) can be used as a silver phytic acid complex. LunarSilver (registered trademark) is as described above.

  Examples of compounds that can be copper ions include metal copper, copper salts, copper complexes, and the like, and copper salts and copper complexes are preferable.

  Metallic copper may be nanoparticulated copper or colloidal copper.

  Examples of the copper salt include copper sulfate, copper chloride, copper oxide, and copper carbonate. Copper sulfate is preferred.

  Copper complexes include copper-ethanolamine, copper-dimethyldithiocarbamate, copper-sulfate, copper-2-ethylhexanoate, copper-quaternary alkylammonium, copper hydroxide (II), basic copper carbonate, copper -Nitrate, copper-8-quinolinolate, copper amino acid complex and the like.

  In the method of the present invention, the proportion of the antibacterial metal (excluding zinc) is usually about 0.00005 (w / v)% or more and about 0.6 (w / v)% or less, preferably about 0.0001 (w / v)% or more and about 0.01 (w / v)% or less, more preferably about 0.0005 (w / v)% or more and about 0.01 (w / v)% or less, and most preferably about 0.005 (w / v)% or more and about 0.01 (w / v)% or less. In the case of a silver salt or a silver complex, the upper limit of addition as silver ions is preferably about 0.01 (w / v)% from the viewpoint of change in appearance of the composition. In the case of a copper salt or a copper complex, the concentration as a copper ion is usually about 0.0001 (w / v)% or more, about 0.5 (w / v)% or less, preferably about 0.0005 (w / v)% or more, About 0.01 (w / v)% or less. From the viewpoint of eye safety, the upper limit of addition as copper ions is preferably about 0.01 (w / v)%.

  In the method of the present invention, various additives such as the tonicity agent, buffering agent, preservatives other than antibacterial metals, stabilizers, antioxidants, chelating agents, pH adjusters, and thickeners are further added. May be used. Details of these various additives are as described above.

  Hereinafter, the present invention will be described more specifically with reference to examples, but it goes without saying that the present invention is not limited thereto.

The storage efficacy of the difluprednate emulsion composition was examined.
1.1 Test Method Staphylococcus aureus (ATCC 6538), Escherichia coli (ATCC 8739) and Pseudomonas aeruginosa (ATCC 9027) were used as test bacteria. Each test bacterium was inoculated on the surface of an agar slant medium and precultured. Pre-culture was performed at 30-35 ° C. for 18-24 hours using Soybean / Casein / Digest agar medium.

The emulsion composition to be used for the test was used as a sample, and the sample was dispensed into 5 sterilized test tubes with stoppers. The test bacteria pre-cultured on the specimen were added and mixed so that the number of bacteria became 10 ⁵ to 10 ⁶ cells / mL, and used as a mixed sample. Each test bacteria was added to the specimen alone. The mixed sample was stored in the dark at 20-25 ° C.

  1 mL was sampled from each mixed sample 6 hours and 24 hours after the start of storage, and the solution was diluted with 9 mL of physiological saline. Further, the same dilution was performed 2-3 times, and 1 mL of each diluted solution was dispensed into a sterile petri dish. Next, 0.1 (w / v)% lecithin · 0.7 (w / v)% polysorbate 80-added soybean / casein / digest agar medium was added to the mixture, followed by culturing at 30 to 35 ° C. for about 3 days. The number of generated colonies was measured, and the number of viable bacteria per 1 mL of the mixed sample was calculated.

1.2 Judgment Criteria If the viable count of all bacterial species decreased by 3 logs or more from the start 24 hours after the start of the test, it was judged to have an immediate effect on storage efficacy and the assessment was relevant. In addition, when immediate effect of further preservation efficacy is required, the viable count of all bacterial species at 24 hours after the start of the test is reduced by 3 logs or more from the start, and all the bacterial species at 6 hours after the start of the test. When the number of viable bacteria decreased by 2 logs or more from the start, it was judged that the storage effect had immediate effect, and the determination was made appropriate.

  The sources of reagents used in the tests are as shown in the table below.

2. Test example 1 (silver compound)
2.1 Preparation Method Preparations having the formulations 1 to 5 shown in Table 2 were prepared. The preparation method was described below.

Preparation method:
100 g of castor oil was weighed into a 200 mL beaker and warmed (water bath temperature: 85-95 ° C.). To this beaker, 1.0 g of difluprednate was added and stirred to dissolve the difluprednate to obtain an oil phase. Separately, 80 g of polysorbate 80 and 44 g of concentrated glycerin were weighed into a 2 L beaker, and 700 mL of water was added. This liquid was heated to about 70 ° C., 1.0 g of sodium acetate hydrate was added with stirring, and the mixture was stirred to obtain an aqueous phase. In Formulation 5, 2.0 g of sorbic acid, 2.0 g of boric acid and 0.4 g of sodium edetate were dissolved in the aqueous phase. The aqueous phase heated to about 70 ° C was stirred (2000 rpm) with a homomixer (TK Robomix, Primix), and the oil phase heated to about 90 ° C was added dropwise to the aqueous phase little by little. The homomixer was rotated at 8000 rpm and stirred at about 70 ° C. for 1 hour. This solution was returned to room temperature, and an appropriate amount of 1 mol / L sodium hydroxide aqueous solution was added to adjust the pH to about 5.5. Purified water was added to make a total volume of 1000 mL to obtain a crude emulsion. Next, using a microfluidizer (M-110EH, Microfluidics Corp.), the crude emulsion was treated for 20 passes at a pressure of about 1500 kgf / cm ² and a temperature of 35 to 45 ° C. to obtain an emulsion stock solution.

  Add 50 mL of the emulsion stock solution to a 100 mL beaker.In Formula 1, 5 mL of 0.16 (w / v)% silver nitrate aqueous solution (0.005 g as silver ions) and in Formula 2, 5 mL of 0.016 (w / v)% silver nitrate aqueous solution (silver 0.0005 g) as ions, or 1 mL of 0.016 (w / v)% aqueous silver nitrate solution (0.0001 g as silver ions) was added for formulation 3. In Formula 4, 1.3 mg of protein silver (0.0001 g as silver ions) was added and dissolved, and in Formula 5, LunarSilver (registered trademark) 0.1 μL (0.00005 g as silver ions) was added. Purified water was added to each solution to make a total volume of 100 mL. The pH of each solution was confirmed to be about 5.5, and the intended emulsion composition was obtained.

2.2 Test Results Table 3 shows the number of viable bacteria in Formulations 1 to 5 24 hours after the start of the test. Formula 1 (silver nitrate, 0.005 (w / v)% as silver ion) and Formula 2 (silver nitrate, 0.0005 (w / v)% as silver ion) met the criteria. However, Formula 3 (silver nitrate, 0.0001 (w / v)% as silver ions) was incompatible with the criteria. Formula 4 (protein silver, 0.0001 (w / v)% as silver ion) and Formula 5 (LunarSilver (registered trademark), 0.00005 (w / v)% as silver ion) met the criteria.

  Table 4 shows the viable cell counts 6 hours after the start of the test in Formulations 1 to 5. Formula 1 (silver nitrate, 0.005 (w / v)% as silver ion) and Formula 5 (LunarSilver (registered trademark), 0.00005 (w / v)% as silver ion) met the criteria. However, Formula 2 (silver nitrate, 0.0005 (w / v)% as silver ions), Formula 3 (silver nitrate, 0.0001 (w / v)% as silver ions), and Formula 4 (protein silver, 0.0001 (w / v) as silver ions) v)%) did not meet the criteria.

3. Test Example 2 (Copper compound)
3.1 Preparation Method Emulsions having the formulations 7 and 8 shown in Table 5 were prepared. The preparation method was described below.

Preparation method:
100 g of castor oil was weighed into a 200 mL beaker and warmed (water bath temperature: 85-95 ° C.). To this beaker, 1.0 g of difluprednate was added and stirred to dissolve the difluprednate to obtain an oil phase. Separately, 80 g of polysorbate and 44 g of concentrated glycerin were weighed into a 2 L beaker, and 700 mL of water was added. While this liquid was heated to about 70 ° C. and stirred, 1.0 g of sodium acetate hydrate was added, and the mixture was stirred to obtain an aqueous phase. The aqueous phase heated to about 70 ° C was stirred (2000 rpm) with a homomixer (TK Robomix, Primix), and the oil phase heated to about 90 ° C was added dropwise to the aqueous phase little by little. The homomixer was rotated at 8000 rpm and stirred at about 70 ° C. for 1 hour. This solution was returned to room temperature, and an appropriate amount of 1 mol / L sodium hydroxide aqueous solution was added to adjust the pH to about 5.5. Purified water was added to make a total volume of 1000 mL to obtain a crude emulsion. Next, using a microfluidizer (M-110EH, Microfluidics Corp.), the crude emulsion was treated for 20 passes at a pressure of about 1500 kgf / cm ² and a temperature of 35 to 45 ° C. to obtain an emulsion stock solution.

  Add 50 mL of the emulsion stock solution to a 100 mL beaker, 0.039 (w / v)% copper sulfate pentahydrate 5 mL (0.0005 g as copper ion) for formulation 7, or 0.039 (w / v)% for formulation 8 1 mL of an aqueous copper sulfate pentahydrate solution (0.0001 g as a copper ion) was added thereto. Purified water was added to each solution to make a total volume of 100 mL. The pH of each solution was confirmed to be about 5.5, and the intended emulsion composition was obtained.

3.2 Test results Table 6 shows the number of viable bacteria in Formulas 7 and 8 24 hours after the start of the test. Formula 7 (0.0005 (w / v)% as copper ion) met the criteria. However, Formula 8 (0.0001 (w / v)% as copper ions) was incompatible with the criteria.

  In order to evaluate the immediate effect of further preservation efficacy, the viable cell count at 6 hours after the start of the test in the formulations 7 and 8 was evaluated (Table 7). Formula 7 (0.0005 (w / v)% as copper ion) met the criteria. However, Formula 8 (0.0001 (w / v)% as copper ions) was incompatible with the criteria.

4. Test Example 3
4.1 Preparation Method As shown in Table 8, emulsion compositions of formulations 9 to 11 containing preservatives other than antibacterial metals were prepared. The preparation method was described below.

Formula 9 preparation method:
50 g of castor oil was weighed into a 100 mL beaker and warmed (water bath temperature: 85-95 ° C.). To this beaker, 0.5 g of difluprednate was added and stirred to dissolve the difluprednate to obtain an oil phase. Separately, 80 g of polysorbate 80 and 22 g of concentrated glycerin were weighed into a 1 L beaker, and 800 mL of water was added. This liquid was heated to about 70 ° C., and while stirring, 0.5 g of sodium acetate hydrate, 1 g of boric acid, 0.2 g of sodium edetate hydrate and 1 g of sorbic acid were added, and the mixture was stirred. The water phase.

The aqueous phase heated to about 70 ° C was stirred (2000 rpm) with a homomixer (TK Robomix, Primix), and the oil phase heated to about 90 ° C was added dropwise to the aqueous phase little by little. The homomixer was rotated at 8000 rpm and stirred at about 70 ° C. for 1 hour. The solution was returned to room temperature, and the pH was adjusted to about 5.5 using a 1 mol / L aqueous sodium hydroxide solution. Purified water was added to this solution to make a total volume of 1000 mL to obtain a crude emulsion. Next, using a microfluidizer (M-110EH, Microfluidics Corp.), the crude emulsion was treated for 20 passes at a pressure of about 1500 kgf / cm ² and a temperature of 35 to 45 ° C. to obtain the desired emulsion composition. .

Formula 10 preparation method:
Formulation 10 was prepared in the same manner as Formulation 9. However, it was prepared by adding 0.1 g of benzalkonium chloride instead of boric acid, sodium edetate and sorbic acid in formula 9.

Formula 11 preparation method:
Formulation 11 was prepared in the same manner as Formulation 9. However, the sorbic acid of Formula 9 was increased to 2 g, and 0.2 g of benzalkonium chloride and 0.1 g of oxyquinoline sulfate were further added.

4.2 Test Results Table 9 shows the viable cell counts 24 hours after the start of the tests for Formulas 9-11. In all formulas 9 to 11, the viable count in all bacterial species did not fall below 10 ² cfu / mL. In addition, Table 10 shows the viable cell counts 6 hours after the start of the tests in prescriptions 9 to 11. In all formulas 9 to 11, the viable count in all bacterial species did not fall below 10 ³ cfu / mL. Therefore, all formulations 9 to 11 did not meet the criteria, and no immediate effect was observed in the storage efficacy.

Summary When sorbic acid, benzalkonium chloride, and oxyquinoline sulfate were added as preservatives, the judgment criteria were not satisfied, and no immediate effect of preservative efficacy was observed.

  On the other hand, by using a silver compound, an immediate effect of storage effect was recognized. When silver nitrate is used, the concentration at which immediate effect of storage effect is recognized is 0.0005 (w / v)% or more as silver ions, and the concentration at which immediate effect is further confirmed is 0.005 (w / v)% as silver ions. That was all. In addition, when LunarSilver (registered trademark) was used, it met the criteria even 6 hours after the start of the test, so if it was added in an amount of 0.00005 (w / v)% or more as silver ions, excellent immediate effect Was considered to be accepted.

  Further, even when a copper compound was used, an immediate effect of storage effect was recognized as in the case of using a silver compound. It was considered that the copper sulfate was added to the emulsion composition in an amount of 0.0005 (w / v)% or more as copper ions, thereby exhibiting an excellent immediate effect of preservation effect.

  Therefore, in the case of the diflupredonate emulsion composition, it has been clarified that an immediate effect of storage effect is recognized by containing an antibacterial metal such as silver or copper (excluding zinc).

  While the invention has been described with emphasis on preferred embodiments, it will be apparent to those skilled in the art that the preferred embodiments can be modified. The present invention contemplates that the present invention may be practiced otherwise than as specifically described herein. Accordingly, this invention includes all modifications encompassed within the spirit and scope of the appended claims.

  The contents of all publications, including the patents and patent application specifications mentioned herein, are hereby incorporated by reference herein to the same extent as if all were expressly cited. .

  This application is based on US Provisional Patent Application No. 61 / 765,349 (filing date: February 15, 2013), the contents of which are hereby incorporated by reference herein in their entirety. Is.

Claims (26)

  An emulsion composition containing difluprednate and an antibacterial metal (excluding zinc).   2. The emulsion composition according to claim 1, wherein the antibacterial metal (excluding zinc) is a salt or complex of the antibacterial metal (excluding zinc).   3. The emulsion composition according to claim 2, wherein the salt or complex of the antibacterial metal (excluding zinc) is at least one salt or complex of the antibacterial metal selected from the group consisting of silver and copper.   4. The emulsion composition according to claim 3, wherein the salt or complex of the antibacterial metal (excluding zinc) is a silver salt or a silver complex.   5. The emulsion composition according to claim 4, wherein the silver salt or silver complex is at least one selected from the group consisting of silver nitrate, protein silver and silver phytic acid complex.   6. The emulsion composition according to claim 4, wherein the concentration of silver salt or silver complex as silver ions is from 0.00005 (w / v)% to 0.6 (w / v)%.   6. The emulsion composition according to claim 4, wherein the concentration of silver salt or silver complex as silver ions is 0.0005 (w / v)% or more and 0.01 (w / v)% or less.   4. The emulsion composition according to claim 3, wherein the salt or complex of the antibacterial metal (excluding zinc) is a copper salt or a copper complex.   9. The emulsion composition according to claim 8, wherein the copper salt or copper complex is copper sulfate.   10. The emulsion composition according to claim 8, wherein the concentration of the copper salt or copper complex as a copper ion is higher than 0.0001 (w / v)% and not higher than 0.5 (w / v)%.   10. The emulsion composition according to claim 8 or 9, wherein the concentration of the copper salt or copper complex as a copper ion is 0.0005 (w / v)% or more and 0.01 (w / v)% or less.   The emulsion composition according to any one of claims 1 to 11, which is an ophthalmic composition.   A method for imparting an immediate effect of storage efficacy to an emulsion composition containing difluprednate, comprising preparing an emulsion composition containing difluprednate and an antibacterial metal (excluding zinc) how to.   14. The method according to claim 13, wherein the immediate effect of the storage effect is to reduce the viable cell count by 3 logs or more within 24 hours after inoculation.   The immediate effect of the storage efficacy is to reduce the number of viable bacteria by 3 logs or more within 24 hours after inoculation and to reduce the number of viable bacteria by 2 logs or more within 6 hours after inoculation. Method.   The method according to any one of claims 13 to 15, wherein the antibacterial metal (excluding zinc) is a salt or complex of the antibacterial metal (excluding zinc).   17. The method according to claim 16, wherein the salt or complex of the antibacterial metal (excluding zinc) is a salt or complex of at least one antibacterial metal selected from the group consisting of silver and copper.   18. The method according to claim 17, wherein the salt or complex of the antibacterial metal (excluding zinc) is a silver salt or a silver complex.   19. The method according to claim 18, wherein the silver salt or silver complex is at least one selected from the group consisting of silver nitrate, protein silver, and silver phytic acid complex.   20. The method according to claim 18 or 19, wherein the concentration of silver salt or silver complex as silver ions is 0.00005 (w / v)% or more and 0.6 (w / v)% or less.   20. The method according to claim 18 or 19, wherein the concentration of silver salt or silver complex as silver ions is 0.0005 (w / v)% or more and 0.01 (w / v)% or less.   18. The method according to claim 17, wherein the salt or complex of the antibacterial metal (excluding zinc) is a copper salt or a copper complex.   23. The method according to claim 22, wherein the copper salt or copper complex is copper sulfate.   The method according to claim 22 or 23, wherein the concentration of the copper salt or the copper complex as a copper ion is higher than 0.0001 (w / v)% and not higher than 0.5 (w / v)%.   The method according to claim 22 or 23, wherein the concentration of the copper salt or copper complex as a copper ion is 0.0005 (w / v)% or more and 0.01 (w / v)% or less.   The method according to any one of claims 13 to 25, wherein the emulsion composition is an ophthalmic composition.